The Cell Analysis Facility (CAF) is a fee for service flow cytometry shared resource operating to provide research flow cytometry services to investigators. The goal of the CAF is to provide comprehensive data collection, data analysis, data interpretation, cell sorting, and education in flow cytometry to all Interested investigators. In addition, the facility provides support for proper sample preparation, as well as education in theory and practice of flow cytometry. The CAF provides state of the art flow cytometry support in all areas of established protocols and continually develops new applications guided by investigator needs. Routine analyses include immunophenotyping, DNA/Cell Cycle determinations, fluorescent protein expression, multiple bead based assays for secreted proteins and cell sorting in bulk or single cell cloning. Given the complexity of the equipment, the CAF personnel assist investigators in the design and interpretation of multi laser, multi parametric experiments. The Cell Analysis Facility, located in a 1200 square foot laboratory in the Durham Research Center I (DRCI) Building, was established in 1980 with the aid of a NIH Shared Instrument Grant and a matching grant from the University of Nebraska Foundation. Current instrumentation in the CAF consists of two (2) 4 laser, 15 color BD Bioscience FACS Aria cell sorters, a 4 laser, 15 color BD Biosciences LSR II, three (3) 2 laser, 4 color BD Biosciences FACS Calibur flow cytometers and a 2 laser, 4 color FACS Array. The 4 color BD Biosciences FACS Calibur flow cytometers have served as the main facility instrument for analysis. The increased demand for sorting prompted the purchase of two (2) Becton Dickinson FACS Aria Cell Sorters and a Miltenyi AutoMACS magnetic bead cell sorter during the past 5 years. Additionally, the CAF provides wet lab space for Investigator or facility initiated sample preparation, incubator space for sample storage prior to and following sorting application, and microscopes (fluorescent and bright field) for viewing cells prior to and following sample preparation activities. In support of these instruments, the CAF provides three workstations with additional copies of Modfit, Flowjo, Cellquest and Diva for post acquisition data analysis and graphic generation for publications and grant submissions. Charles Kuszynski, Ph.D. has been the facility director for the Cell Analysis Facility for 30 years. He has significant experience in developing protocols and analyzing data, and he was trained by Becton Dickinson as a field engineer to maintain the instrumentation in the laboratory.